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A new species, Mazosia hainanensis, is described from Hainan, China. It is most similar to M. pseudobambusae but differs by its yellowish green thallus and smaller ascospores (14–18.5 × 3–4 μm). An analysis of its relationships based on molecular phylogeny is given. A key to the Mazosia species recorded in China is also presented.

LncRNA PVT1 has been implicated in numerous pathophysiological processes and diseases, especially cancers. However, the role and mechanism of PVT1 in the tumorigenesis of glioblastoma remain unclear. We investigated the alteration of PVT1 and its key functions in glioblastoma. PVT1 was upregulated and associated with poor prognosis in glioblastoma. We demonstrated that PVT1 silencing suppressed cell proliferation, colony formation, and orthotopic xenograft tumor growth. Mechanistic investigations found that PVT1 interacted with TRIM24 directly and increased its protein stability. PVT1 recruited COPS5 to deubiquitinate TRIM24; reciprocally, PVT1 depletion impaired the interaction between COPS5 and TRIM24, resulting in decreased expression of TRIM24. PVT1, TRIM24, and COPS5 coordinately contributed to the activation of STAT3 signaling and malignant phenotype of glioblastoma. Collectively, this study elucidates the essential role of PVT1 in the tumorigenesis of glioblastoma, which provides candidacy therapeutic target for glioblastoma treatment., Graphical Abstract, PVT1 was highly elevated in glioblastoma (GBM) patients and associated with poor prognosis. PVT1 recruited COPS5 and bound TRIM24 to deubiquitinate and stabilize TRIM24. PVT1 contributed to the tumorigenesis of GBM via forming a complex with TRIM24 and COPS5 and activating STAT3 signaling.

The antihypertensive activity of yeast hydrolysate (YH) was confirmed in our previous study. However, the critical peptides in YH and the underlying mechanisms have not been fully elucidated. This study aimed to explore the angiotensin-converting enzyme (ACE) inhibitory peptides in YH and illustrate their molecular and cellular mechanisms. The potential of YH-derived peptides was evaluated by in silico methods, followed by in vitro verification. A new competitive ACE inhibitory peptide, VIPVPFF (V7), with an IC50 value of 10.27 μM, was screened. YH and V7 increased the nitric oxide (NO) levels, upregulated GUCY1A1 gene expression (approximately 15-fold), and functioned in several hypertension-related pathways in human umbilical vein endothelial cells (HUVECs). This study revealed the antihypertensive mechanisms of YH and V7, laying down a theoretical basis for their application.

Trapelia calyciformis is described from China as a new species. The lichen is characterized by its gray-white thallus, zeorine apothecium with black disc and cracked excipulum, narrow paraphyses branched near the tip, 8-spored asci, aseptate ascospores with one vacuole, and the presence of gyrophoric acid. The specimens examined were deposited in LCUF. A key to the species of Trapelia reported in China is presented.

Gastric cancer (GC), as one of the most prevalent malignancies, contributes to the high morbidity and mortality worldwide. By analyzing the bioinformatics, qRT-PCR and IHC assays, we found that CLEC5A is overexpressed in GC and associated with poorer prognosis. CLEC5A silencing inhibits cell growth and DNA replication and induces cell cycle arrest and cell apoptosis. Bioinformatics analyses and Western blotting revealed that CLEC5A depletion led to the dysregulation of the PI3K/AKT/mTOR pathway. CLEC5A-mediated GC proliferation and anti-apoptosis were impaired by blocking the PI3K/AKT/mTOR pathway with LY294002. We hypothesize that CLEC5A is of vital importance to GC initiation and progression via the PI3K/AKT/mTOR pathway, and that our results might represent promising therapeutic strategies for GC patients.

The changes in the gluten network during extrusion treatment were studied by assessing the polymerization behavior of glutenin. Gluten samples were extruded at different barrel temperatures, screw speeds, and flow rates. The results indicated that high molecular weight glutenin subunits increased while free sulfhydryl groups and low molecular weight glutenin subunits decreased as the screw speeds and flow rates increased during extrusion treatment. Specific β-sheet structures of gluten clearly increased, while α-helices and β-turns fluctuated during extrusion processing, thus forming a tight gluten network. The characteristics of the protein network were evaluated by confocal laser scanning microscopy. The results showed that a homogeneous and denser gluten network was formed at higher extrusion temperatures during the extrusion process, which may be related to the polymerization of low-molecular-weight glutenin subunits. This study provides a theoretical basis for the improvement and regulation of extrusion quality during the gluten extrusion process.

The aim of this study was to analyze the physicochemical and structural characteristics of the Venn components of wheat gliadin to provide theoretical basis of gliadin for processing in dough and Chinese steamed bread. Eight Venn components, Gli-8, Gli-9, Gli-10, Gli-11, Gli-12, Gli-13, Gli-14, and Gli-15, were extracted from wheat gliadin based on their solubility. The results of physicochemical characteristics showed that the differences in the contents, TDS, electrical conductivity, particle size and zeta potential of Venn components were significant, respectively. The content of Gli-15 in gliadin was the highest, and the content of Gli-9 was the lowest. The TDS value of Gli-9 was the highest (336.0), and the TDS value of Gli-15 was the lowest (52.0). The electrical conductivity of Gli-9 was the highest, which was 7.54 times the lowest value of Gli-11. The zeta potential of Gli-9 was −25.2 mV, and the zeta potential of the Gli-15 was −7.64 mV. However, the difference in the pH values was not significant. The results of UV spectrum and FTIR analysis showed that the secondary structures of the Venn components had significant differences. The results of the XRD patterns indicated that the Venn components might not be a single substance. The results of CLSM images implied that the molecular interactions among the components were varied. Hence, the results could provide research materials and basic data for deep processing and utilization of gliadin. Keywords: Wheat gliadin, Venn components, Physicochemical properties, Structural characteristics

Background Circular RNAs (circRNAs) are recently identified as gene regulators in mammals and play important roles in carcinogenesis of cancer. For example, circRNA_PTN has been recognized as a biomarker of human cancer and is overexpressed in glioma. The molecular function of circRNA_PTN and its downstream targets in glioma, however, remains elusive. Methods Quantitative polymerase chain reaction analysis was used to measure the expression of circular RNA pleiotrophin (circ_PTN) and miR-122. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, propidium iodide and Annexin-V/propidium iodide assay were performed to determine cell proliferation and apoptosis of glioma cells. Circular RNA Interactome and TargetScan were used to predict the potential microRNA targeting of circ_PTN and the potential targets of miR-122, respectively. Luciferase activity assay was used to validate these interactions. Downstream molecular mechanisms, including SRY-box transcription factor 6 (SOX6), extracellular regulated protein kinases (ERK), Cyclin D1, B-cell lymphoma-2 (BCL-2) and BCL2 associated X, apoptosis regulator (BAX), were determined by western blot. Results Circ_PTN was overexpressed in glioma cells, and its knockdown induced cell proliferation inhibition, cell cycle arrest and apoptosis in glioma cells. The target microRNA of circ_PTN was predicted to be miR-122, the expression of which was negatively correlated with circ_PTN in glioma cells. Moreover, SOX6 was predicted as a potential target of miR-122, and miR-122 overexpression decreased SOX6 expression. MiR-122 inhibitor reversed the tumor-suppressing effects of circ_PTN knockdown, while overexpression of SOX6 impaired the miR-122 overexpression-induced cell growth inhibition and apoptosis. In addition, mitogen activated kinase-like protein (MAPK)/ERK pathway was involved in circ_PTN/miR-122/SOX6 axis. Conclusions Circ_PTN acted as a sponge of miR-122 and upregulated miR-122 target SOX6, thus promoting carcinogenesis of glioma cells.

Background: Circular RNAs (circRNAs) are widely expressed non-coding RNAs in eukaryotic cells, involved in regulating tumorigenesis of several types of cancers. However, the expression profiles and the precise functional role in glioblastoma remain unclear. Results: Circ-EPB41L5 was downregulated in glioblastoma tissues and cell lines compared to the normal brain tissues and cell lines. Low circ-EPB41L5 expression was correlated to the poor prognosis of glioblastoma patients, while the overexpression inhibited proliferation, clone formation, migration, and invasion abilities of glioma cells, and the suppression had counter effects. Furthermore, RNA-seq results determined that the host gene was the target gene of circ-EPB41L5, which served as a sponge against miR-19a and inhibited miR-19a activity from upregulating the expression of EPB41L5. Finally, we found that circ-EPB41L5 regulated the RhoC expression and phosphorylation of AKT through EPB41L5. Conclusion: The current study highlights a novel suppressive function of circ-EPB41L5 and reveals that circ-EPB41L5/miR-19a/EPB41L5/p-AKT regulatory axis plays a striking role in the progression of glioblastoma, which provides a novel insight into the mechanisms underlying glioblastoma. Methods: The expression profiles of circRNAs in glioblastoma were determined by Illumina HiSeq from six glioblastoma tissues and six normal brain tissues. Then, the correlation between circ-EPB41L5 expression and clinical features and the survival time of 45 glioblastoma patients was detected. The interaction between circ-EPB41L5, miR-19a, and EPB41L5 was assessed by luciferase reporter and RNA pull-down assays. The effects of expression of the ectopic intervention of circ-EPB41L5 or EPB41L5 on proliferation, clone formation, migration, and invasion in vitro and tumorigenesis in vivo were observed to evaluate the function of circ-EPB41L5 or EPB41L5.

The present study was aimed to investigate change in physicochemical characteristics and glutenin macropolymer (GMP) molecular weight distribution during postharvest wheat maturation. The freshly harvested wheat was stored under four different conditions (WT1, WT2, WT3 and WT4) for specified times. During maturation, the strengthening of wheat gluten structure was observed by increase in gluten index, swelling index of glutenin and sodium dodecyl sulphate (SDS) sedimentation value. The postharvest maturation resulted in contents increase of glutenin, glutenin/gliadin ratio, and GMP, and the decline of gliadin content. The contents of free sulfhydryl in GMP reduced, and S-S content rose during the whole maturation. Visible aggregates of GMP were observed in SDS-PAGE patterns for WT3 and WT4 samples after 4 weeks of storage. Moreover, in these two samples, polymerisation of gluten proteins into GMP was observed in size-exclusion high performance liquid chromatography patterns during storage. Some low molecular weight glutenin subunits (LMW-GS) incorporated into the aggregate, or combination of high molecular weight glutenin subunits and LMW-GS formed large polymeric glutenins by S-S linkage, and then assembled into GMP in postharvest wheat maturation. GMP polymerisation probably results in the improvement of wheat quality.

Objective. To determine the subcellular localization of epithelial cell adhesion molecule (EpCAM) in labial salivary gland (LSG) and evaluate the diagnostic use of the extracellular domain of EpCAM (EpEX) and intracellular domain (EpICD) for primary Sjögren’s syndrome (pSS). Methods. Immunohistochemical (IHC) analysis was conducted using EpEX and EpICD domain-specific antibodies on labial salivary gland biopsy (LSGB) from participants. Chi-square or Fisher’s exact analysis, Mann–Whitney U-test, and Kruskal-Wallis test compared differences among groups. Independent risk factors of pSS were determined by multiple logistic regression analysis. Receiver-operator characteristic curves (ROC) were carried out to estimate the diagnostic value. Results. Compared to non-SS controls, loss of membranous EpEX and EpICD expression was observed in LSGB of pSS patients, which occurred in parallel with increased accumulation of cytoplastic and nuclear EpICD. The subcellular EpEX/EpICD expressions were associated with various features of pSS patients, especially histopathological grade of LSGB. Furthermore, high IHC scores of membranous EpEX were independent risk factors for pSS, even for the pSS patients at early stage. The IHC scores of subcellular EpEX/EpICD were of great diagnostic value for pSS with high sensitivity (70-80%) and specificity (85-95%). Conclusion. This study first found the aberrant expression pattern of EpCAM in LSG of pSS patients. The IHC scores of subcellular EpEX/EpICD were demonstrated to have the potential to act as diagnostic biomarkers for pSS.

In this work, the fermentation conditions and the antibacterial characteristics of Eucommia ulmoides leaves vinegar (EV) were studied. By single factor orthogonal test, it was found that under optimal fermentation conditions (bran addition 10%, sugar addition 8%, leaven addition 0.3% and acetic acid bacteria solution 12%), the acetic acid content and CA content of EV were 45.5 ± 2.8 mg/mL and 0.98 ± 0.08 mg/mL, respectively. Then, by the disc diffusion method, it was concluded that the antibacterial effect of EV was significantly higher than that of Eucommia ulmoides leaves enzymatic hydrolysate and Zhenjiang aromatic vinegar (P

Melatonin (MEL) has been demonstrated to exert a protective effect against subarachnoid hemorrhage (SAH), and nitric oxide (NO) has been shown to play an important role in the pathogenesis of vasospasm. This study aims to explore the underlying molecular mechanisms of MEL in the control of vasospasm following SAH. MEL administration attenuates SAH-induced vasospasm and neurobehavioral deficits. Expressions of H19, eNOS, and miR-675 are low in the SAH group, while expressions of miR-138 and HIF1α are high in the SAH group. Also, MEL treatment upon SAH rats completely restores the dysregulation of H19, eNOS, miR-675, miR-138, and HIF1α to their normal levels. Moreover, MEL dose dependently increases the luciferase activity of H19 promoter and hence the expression of H19. Additionally, H19 directly targets miR-675 and miR-138 to increase miR-675 expression and inhibit miR-138 expression. As virtual target genes of miR-675 and miR-138, respectively, HIF1α and eNOS are also regulated by the treatment with MEL. In particular, MEL treatment increases the expression of miR-675 and eNOS level while decreasing the expression of miR-138 and HIF1α in a dose dependent manner. Our study found that MEL ameliorates post-SAH vasospasm by regulating the expression of eNOS and HIF1α via the H19/miR-138/eNOS/NO and H19/miR-675/HIF1α signaling pathways.

A new species, Coenogonium hainanense, is proposed, characterized by its small slightly transparent pale yellow to brownish apothecia with plane to concave discs; slightly thick, not prominent, denticulate margins with short dense hairs; and uniseriate or irregularly biseriate, ellipsoid 1-septate ascospores (10–11.25 × 2.5–3.75 μm). Eight additional Coenogonium species reported as new to China are C. coronatum, C. curvulum, C. fallaciosum, C. geralense, C. hypophyllum, C. moniliforme, C. siquirrense, and C. usambarense. Descriptions and distributions of the new species and records are presented, and a key to the 20 Coenogonium species known from China is provided.

Background and Purpose: Sirt5 (Sirtuin 5) desuccinylates multiple metabolic enzymes and plays an important role in maintaining energy homeostasis. The goal of this study was to determine whether Sirt5-mediated desuccinylation restores the energy metabolism and protects brain against subarachnoid hemorrhage (SAH). Methods: Male C57BL/6 or Sirt5 −/− mice were used. The endovascular perforation SAH model was applied. Protein lysine succinylation in the brain cortex was examined using liquid chromatography-tandem mass spectrometry analysis. The brain metabolism was evaluated by measurement of brain pH as well as ATP and reactive oxygen species level. Neuronal cell death and neurobehavioral deficits were assessed 24 hours after SAH. The expression and desuccinylation activity of Sirt5, lysine succinylation of citrate synthase and ATP synthase subunits were investigated by Western blot, immunohistochemistry, and ELISA in SAH mice and patients. Furthermore, the benefits of resveratrol-mediated Sirt5 activation were investigated. Results: A total of 211 lysine succinylation sites were differentially expressed on 170 proteins in mice brain after SAH. Thirty-nine percent of these succinylated proteins were localized in mitochondria and they are related to energy metabolism. SAH caused a decrease of Sirt5 expression and succinylated citrate synthase as well as the subunits of ATP synthase, subsequently lowered brain pH, reduced ATP and increased reactive oxygen species production, leading to neuronal cell death, and neurological deficits. Knockdown of Sirt5 aggravated SAH-induced effects, mentioned above. Administration of resveratrol resulted in activation of Sirt5. The activation was accompanied both with restoration of the mitochondrial metabolism and alleviation of early brain injury as well as with desuccinylating citrate synthase and ATP synthase. Conclusions: Protein lysine succinylation is a biochemical hallmark of metabolic crisis after SAH, and disruption of lysine succinylation through activation of Sirt5 might be a promising therapeutic strategy for the treatment of SAH.

During a study of the genus Mazosia in China, a new species, named here M. weii, was found. Its position was found to be supported by molecular data of the mtSSU and nuLSU markers. The new species is characterized by a pilose thallus with brown verrucae and 3-septate ascospores measuring 22.5–35 × 4–6.25 µm. The morphological, anatomical and chemical characteristics are described in detail and a key to Mazosia with pilose thalli is added.

Butelase-1 is an efficient ligase from Clitoria ternatea with wide applications in the food and biopharmaceutical fields. This research aimed to achieve high-efficiency expression of butelase-1 and explore its application in food-derived angiotensin I-converting enzyme (ACE) inhibitory peptides. The recombinant butelase-1 zymogen was prepared at a yield of 100 mg/L in Escherichia coli and successfully activated at pH 4.5, resulting in a 6973.8 U/L yield of activated butelase-1 with a specific activity of 348.69 U/mg and a catalytic efficiency of 9956 M-1 s-1. Activated butelase-1 exhibited considerable resistance to Tween-20, Triton X-100, and methanol. The "traceless" cyclization of ACE inhibitory peptides was realized using activated butelase-1, which resulted in higher stability and ACE inhibitory activity than those of the linear peptides. Our work proposed an efficient method for the preparation of butelase-1 and provided a promising model for its application in food fields.

In this study, we revealed the differential proteins from the wheat endosperms using proteomic analysis and investigated their surface properties. The pattern of the polypeptides obtained from the Yangmai-15 and Yangmai-16 wheat varieties were compared using two-dimensional polyacrylamide gels. In addition, we compared the characteristics of the grain such as grain hardness, protein content, wet gluten, dough development time, dough stability, gliadin and glutenin contents between Yangmai-15 and Yangmai-16, and the results were significantly different. Notably, 216 and 197 protein spots were separated from Yangmai-15 and Yangmai-16, respectively. The isoelectric points of the identified proteins ranged from 4 to 10 and the molecular weights of proteins varied from 10 to 100 kDa. Further, 21 and 8 specific differential protein spots were identified from the flour of Yangmai-15 and Yangmai-16, respectively. The surface properties of identified peptides consisted of hydrophobic or hydrophilic residues, as well as randomly scattered residues. The proteomic analysis of the wheat endosperms provides a novel insight into the biochemical basis for the differences in physicochemical properties between the soft and hard wheat varieties. Keywords: Soft and hard wheat, Proteome, Peptides surfaces, Endosperm proteins

Tripartite motif protein 32 (TRIM32), an E3 ubiquitin ligase, has been reported to participate in many human cancers. However, the underlying role of TRIM32 in glioma remains largely unknown. Here, we aimed to explore the function of TRIM32 in glioma cells and the clinical implications and found that TRIM32 was upregulated in glioma tissues. Consistently, overexpression of TRIM32 promoted glioma U87 and U251 cell proliferation and conferred cell resistance to temozolomide (TMZ). Conversely, knockdown of TRIM32 inhibited glioma cells proliferation in vitro and in vivo and sensitized glioma cells to the treatment of TMZ in a p53-dependent and -independent manner. Mechanistically, knockdown of TRIM32 induced apoptosis of U87 an U251 cells. In addition, TRIM32 interacted with the antiapoptotic proteins BCL-xL and BCL-w, which antagonized the inhibitory effect of TRIM32 knockdown in U87 cells. Together, our study uncovered the role of TRIM32 in glioma and TRIM32 may be a potential therapeutic target for gliomas.

BackgroundRegenerative medicine and tissue engineering have brought new therapeutic prospects to the treatment of soft tissue defects, but the selection of seed cells is the key to treatment. Adipose-derived stem cells (ASCs) have always been a popular candidate for seed cells because of their rich sources, easy access, high plasticity, and strong value-added capabilities. The purpose of the current study is to explore the role of PACAP -derived peptide MPAPO on the adipogenic differentiation of ASCs and its molecular mechanism.MethodsThe effect of MPAPO on the proliferation of adipose-derived stem cells were detected by CCK-8 assay and PI single-staining-flow. To reveal the direct effect of MPAPO on the adipogenic differentiation of ASCs, a model of adipogenic differentiation of adipose stem cells was established. In addition, adipogenic differentiation capacity was assessed using Oil-Red-O Staining, Triglyceride (TG) assay and quantification of gene expression. Finally, the relationship between ASCs adipogenic differentiation and the ERK signaling pathway was explored by Western blot.ResultsMPAPO treatment can significantly promote the proliferation of ASCs. In addition, PACAP treatment improves the adipogenic differentiation efficiency of ASCs, including promoting the accumulation of lipid droplets and triglycerides, and the expression of adipogenic-related transcription factors PPARγ and C/EBPα. The mechanism studies showed that MPAPO selectively binds to the PAC1 receptor to promote the adipogenic differentiation of ASCs via activating the ERK signaling pathway.ConclusionsThe present study shows that MPAPO could promote the adipogenic differentiation of ASCs by activating the ERK signaling pathway, and provide relevant experimental evidence for the filling of clinical tissue defects.

The influence of extrusion temperature on protein components and aggregation of wheat gluten (WG) and wheat gluten-peanut oil complexes (WPE) during extrusion with the addition of peanut oil was studied. Gliadin content and wheat gluten extractability decreased and glutenin content increased as extrusion temperature increased. At the same extrusion temperature, the gliadin content in WPE was higher than that in WG. The addition of peanut oil also resulted in the higher gluten extractability of WPE than WG. Increasing extrusion temperature also increased the average molecular weight of glutenin and gliadin. The decreased free sulfhydryl (SH) and increased disulfide bonds (SS) indicated that wheat gluten aggregation was promoted, via disulfide cross-linking, when extrusion temperature increased. Furthermore, increased temperature promoted the aggregation of gluten by increasing sulfhydryl-disulfide bond (SH-SS) interchange during extrusion. When the secondary structure of wheat gluten was analyzed by circular dichroism, the relative gluten α-helix content was decreased and the relative β-sheet content was increased. Also, the results of scanning electron microscopy (SEM) showed the size of the resultant particles increased with temperature, and the mean particle size of WPE was higher than WG. This research shows that extrusion temperature promotes gluten aggregation of WG and WPE. It provides basic data to support the study of gluten-lipid extrusion in the field of protein processing.

The rapid development of transcriptome sequencing technology has contributed to the discovery of numerous genes in plant; however, the role of gene expression in postharvest wheat remains largely unexplored. In this study, differentially expressed genes (DEGs) were identified by RNA‐seq in different quality wheats. The 102.6 Gb clean reads had been yielded from the nine RNA‐seq libraries. Typically, there were 1791 upregulated and 2,677 downregulated DEGs, respectively, in strong‐gluten wheat compared with weak‐gluten wheat. Specifically, a total of 4,468 DEGs were classified into 286 Gene Ontology (GO) terms and 131 Kyoto Encyclopedia of Genes and Genomes terms (KEGG). Moreover, the storage protein components, starch and sucrose metabolism, and plant hormone signal transduction‐related genes were discovered, which had involved 109 DEGs. The wet gluten proteins content was 35.24% and 17.36%, and the glutenin macropolymer content was 6.38% and 5.01% between the strong‐ and weak‐gluten wheat, respectively. The POD activities of the different quality wheats were 6,571.14, 5,341.24, and 4,851.48 U/g/min, respectively. The significant difference of starch and sucrose metabolism, hormone, POD, and CAT enzyme along with the higher ATPase activity might potentially affect gluten polymerization, which might thereby result in the different qualities of wheats., This work firstly subjected transcriptome to reveal the effect of the different quality wheat, and we found that the formation of different quality wheats may be due to the difference of GMP, storage protein, hormone, protein isomerase, antioxidase activity, and energy metabolism.

In the pathogenic fungus Candida albicans, phosphoregulation of the checkpoint kinase Rad53 plays a crucial role in the filamentous growth response to genotoxic stresses. The protein phosphatase 4 (PP4) complex, containing Pph3 and either Psy2 or Psy4, is proved to play a critical role in Rad53 dephosphorylation. In previous studies, we characterized CaPtc2 (the ortholog of both Ptc2 and Ptc3 in Saccharomyces cerevisiae) as a potential DNA-damage-related protein phosphatase. In this study, we checked the genetic interaction of PTC2 with the PP4 complex in the DNA damage response pathway. The results suggest that Ptc2 shows a negative genetic interaction with Pph3, but positive genetic interaction with either Psy2 or Psy4 in response to genotoxic stress. Deletion of PTC2 alone resulted in no significant change in cell virulence, but double deletion of PTC2 PPH3 significantly decreased virulence, while double deletions of either PTC2 PSY2 or PTC2 PSY4 caused virulence levels similar to that shown by PSY2 or PSY4 single-gene deletion cells. Taken together, we propose that Ptc2 in C. albicans plays a compensatory role for Pph3 but is dependent on Psy2 and Psy4 in regulation of DNA damage and cell virulence.

Wheat post-harvest maturation induced baking and technological quality improvement through a series of biochemical and colloidal changes. Weak-, middle-, and strong-gluten wheat displayed varying gluten network structures that determined the flour ingredient formulations and processing conditions. However, the aggregation and structural properties of wheat with different gluten strengths post-harvest remain largely unexplored. In this study, we investigated changes in the aggregative properties of gluten protein, gluten composition, S–S content, network structure, and secondary structures of weak-, middle-, and strong-gluten wheat during post-harvest maturation. The results indicated that the macromolecular aggregation of gluten proteins was impaired in weak-gluten wheat, while it was enhanced for middle- and strong-gluten wheat during storage. Post-harvest maturation resulted in an increase in glutenin content and a decline in the gliadin and gliadin/glutenin ratio in middle- and strong-gluten wheat as well as a decreased glutenin content in weak-gluten wheat. Moreover, additional gluten subunits were observed in middle- and strong-gluten wheat, but no substantial change was observed in weak-gluten wheat with long storage times. The disulfide bond content of gluten protein for middle-gluten and strong-gluten gradually increased but declined for weak-gluten wheat. Secondary structure analysis of gluten indicated that post-harvest maturation caused the conversion of α-helix to random coil for weak-gluten wheat, β-turn and random coil to α-helix for middle-gluten wheat, and β-turns to α-helix for strong-gluten wheat, which led to a disordered structure for weak gluten and an ordered stable gluten network for middle- and strong-gluten. Thus, the increased S–S and α-helix content induced by post-harvest maturation enhanced the aggregation of gluten proteins for middle- and strong-gluten wheat, resulting in a denser network structure. Conversely, the decrease in the content of α-helix resulted in the existence of a looser gluten network structure for weak-gluten wheat during post-harvest maturation.

Aldehydes are effective crosslinkers for fabricating protein-based films. Herein, dialdehyde polysaccharides (DPs) were synthesized and applied to prepare the functional gliadin films cooperated by citric acid. The physicochemical properties of the gliadin film-forming solutions and functional properties of the films were fully explored, as well as the formation mechanisms of the films were investigated. Results showed that all the DPs crosslinked gliadin films had a transparent and ultraviolet resistance property. And the mechanical properties, water-resistant properties, thermal stability, antibacterial properties of the gliadin films were all advanced by DPs, in which dialdehyde dextrin crosslinked gliadin films (DAD-film) showed the best performances. And the Schiff base bonds, hydrophobic interactions, and hydrogen bonds were the main molecular interactions in the DPs crosslinked gliadin films, which worked together to achieve the related properties of the films. Therefore, the DPs, especially DAD, could be a potential crosslinker to fabricate the gliadin-based films with functional properties. And this research will highlight the new strategies for the usage of gliadin or other cereal prolamins for packaging.

Gliadin is an ideal matrix for fabricating biodegradable bioplastic films. However, the fabricating of gliadin-based films with functional properties is still a challenge for many fields. Here, a series of gliadin-based films were prepared by crosslinking gliadin with cinnamaldehyde (CA) at different pH (pH 6.5, 7, 8, 9, & 10). Subsequently, the mechanical properties, water-resistant properties, thermal properties, antibacterial properties, and formation mechanisms of the films were systematically investigated. All the CA crosslinked films presented transparency and resistance to ultraviolet (UV) radiation. Significant increase in tensile strength (24-fold), elongation (>477%), water-resistant properties, and long-term water stability (>120 h), implying the highly crosslinking was formed in the CA crosslinked films (Schiff base, Michael's addition, and hydrogen bonds). Meanwhile, the tea polyphenols-loading properties and thermodynamic stability of the films were also enhanced by CA crosslinking under alkaline conditions. Moreover, CA crosslinked films performed splendid antimicrobial activities. The present study indicated that the overall properties of the gliadin-based films can be enhanced by the addition of CA under alkaline conditions. These results may be useful in the development of new applications for gliadin or other cereal prolamins, like drug delivery or packaging materials.

AIM To clarify the mechanisms of HOX transcript antisense intergenic RNA (HOTAIR) in gastric cancer (GC) migration and invasion. METHODS Quantitative real-time polymerase chain reaction (qPCR) was used to detect the expression level of HOTAIR in GC tissues. The correlation of its expression with clinicopathological features was analyzed. Area under receiver operating characteristic curve (AUCROC) was constructed to evaluate the diagnostic value of HOTAIR. Wound-healing assay and Transwell assay were performed to detect the biological effects of HOTAIR in GC cells. qPCR, western blot and immunohistochemistry were used to evaluate the mRNA and protein expression of E-cadherin. RNA-binding protein immunoprecipitation was used for the analysis of EZH2 interactions with HOTAIR. Chromatin immunoprecipitation assay was performed to investigate direct interactions between EZH2 and E-cadherin. RESULTS The expression of HOTAIR was up-regulated in GC tumorous tissues compared with the para-tumorous tissues (P < 0.001). Its over-expression was correlated with tumor-node-metastasis (TNM) stage (P = 0.024), tumor invasion (P = 0.018), lymph node metastasis (P = 0.023), and poor prognosis (P < 0.001). Multivariate Cox regression analysis confirmed expression of HOTAIR as an independent predictor of overall survival (P = 0.033), together with TNM stage (P = 0.002) and lymph node metastasis (P = 0.002). The AUCROC was up to 0.709 (95%CI: 0.623-0.785, P < 0.001). Knockdown of HOTAIR by siRNA in GC cells suppressed the migration and invasion of GC cells. Significantly negative correlation between HOTAIR and E-cadherin was found in GC tissues and cell lines, and HOTAIR contributed to the regulation of E-cadherin through binding to EZH2 with the E-cadherin promoter. CONCLUSION HOTAIR may play a pivotal role in tumor cell migration and invasion. It can be used as a potential diagnostic and prognostic biomarker for GC.

In the framework of continuing studies on the Graphidaceae in China, the status of all taxa traditionally assigned to the genus Graphina reported from China are resolved in the present paper. Five new combinations are made, namely Diorygma isabellinum (Zahlbr.) Z.F. Jia & Lücking, comb. nov., Fissurina adscribens (Nyl.) Z.F. Jia & Lücking, comb. nov., Graphis lecanactiformis (Zahlbr.) Z.F. Jia & Lücking, comb. nov., Phaeographis haloniata (Zahlbr.) Z.F. Jia & Lücking, comb. nov. and Platygramme taiwanensis (J.C. Wei) Z.F. Jia & Lucking, comb. nov. Five new synonymies were found: Graphina olivascens Zahlbr. (= Fissurina adscribens), Graphina plumbicolor Zahlbr. (= Phaeographis haloniata), Graphina roridula Zahlbr. and its variety platypoda Zahlbr. [= Diorygma pachygraphum (Nyl.) Kalb, Staiger & Elix], and Graphina taiwanensis f. obscurata J.C. Wei (= Platygramme taiwanensis).

Inorganic phosphate (Pi) starvation severely affects the normal growth and development of plants. Here, a Pi-responsive gene, named MdMYB2 (MDP0000823458), was cloned and functionally identified in apple. Overexpression of MdMYB2 regulated the expression of Pi starvation-induced (PSI) genes and then promoted phosphate assimilation and utilization. The ectopic expression of MdMYB2 in Arabidopsis influenced plant growth and flowering, which was partially rescued by application of exogenous gibberellin (GA). These results indicated that MdMYB2 may be an essential regulator in phosphate utilization and GA-regulated plant growth and development.

Biocatalytic degradation technology has received a great deal of attention in water treatment because of its advantages of high efficiency, environmental friendliness, and no secondary pollution. Herein, for the first time, horseradish peroxidase and mediator syringaldehyde were co-immobilized into functionalized calcium alginate composite beads grafted with glycidyl methacrylate and dopamine. The resultant biocatalyst of the co-immobilized horseradish peroxidase-syringaldehyde system has displayed excellent catalytic performance to degrade indole in water. The degradation rate of 100% was achieved in the presence of hydrogen peroxide even if the indole concentration was changing from 25 mg/L to 500 mg/L. If only the free enzyme was used under the identical water treatment conditions, the degradation of indole could hardly be observed even when the concentration of indole is low at 25 mg/L. This was attributed to the effective co-immobilization of the enzyme and the mediator so that the catalytic activity of horseradish peroxidase and the synergistic catalytic action of syringaldehyde could be fully developed. Furthermore, while the spherical catalyst was operated in succession and reused for four cycles in 50 mg/L indole solution, the degradation rate remained 91.8% due to its considerable reusability. This research demonstrated and provided a novel biocatalytic approach to degrade indole in water by the co-immobilized horseradish peroxidase-syringaldehyde system as biocatalyst.

The incidence of Lyme disease has continued to rise despite attempts to control its spread. Vaccination of zoonotic reservoirs of human pathogens has been successfully used to decrease the incidence of rabies in raccoons and foxes. We have previously reported on the efficacy of a vaccinia virus vectored vaccine to reduce carriage of Borrelia burgdorferi in reservoir mice and ticks. One potential drawback to vaccinia virus vectored vaccines is the risk of accidental infection of humans. To reduce this risk, we developed a process to encapsulate vaccinia virus with a pH-sensitive polymer that inactivates the virus until it is ingested and dissolved by stomach acids. We demonstrate that the vaccine is inactive both in vitro and in vivo until it is released from the polymer. Once released from the polymer by contact with an acidic pH solution, the virus regains infectivity. Vaccination with coated vaccinia virus confers protection against B. burgdorferi infection and reduction in acquisition of the pathogen by naïve feeding ticks.

Spontaneous axonal regeneration of neurons does not occur after spinal cord injury because of inhibition by myelin and other inhibitory factors. Studies have demonstrated that blocking the Rho/Rho-kinase (ROCK) pathway can promote neurite outgrowth in spinal cord injury models. In the present study, we investigated neurite outgrowth and neuronal differentiation in neural stem cells from the mouse subventricular zone after inhibition of ROCK in vitro. Inhibition of ROCK with Y-27632 increased neurite length, enhanced neuronal differentiation, and upregulated the expression of two major signaling pathway effectors, phospho-Akt and phospho-mitogen-activated protein kinase, and the Hippo pathway effector YAP. These results suggest that inhibition of ROCK mediates neurite outgrowth in neural stem cells by activating the Hippo signaling pathway.

Fabricating a hydrogel with high strength and toughness is still a challenge in many fields. Here, we prepared gliadin-based hydrogels by chemical cross-linking gliadin in acetic acid solution (GS) with glutaraldehyde (GA). Subsequently, the overall properties of the fabricated hydrogels were systematically investigated in terms of their mechanical properties, swelling ratio, weight loss, thermal stability, and the chemical/physical interactions in hydrogels. Results showed that the gliadin-based chemically cross-linked hydrogels exhibited excellent mechanical properties. The optimized hydrogel exhibited the compressive stress of 1.8 MPa at a strain of 70%, and an excellent self-recovery property after 30 cycles of loading-unloading treatments. The strength and toughness of the hydrogels could be tailored by adjusting the ratio of GS/GA. The chemical cross-linking (aldehyde-ammonia reaction) was the main molecular interaction in the hydrogels, including single-/multi-site crosslinking, and the hydrogen bond was the only physical cross-linking in the hydrogels. Moreover, the swelling ratio of the fabricated hydrogels performed a concentration negative-dependency in GA or GS concentration. And a higher GS concentration (40%) with an appropriate GA content (3.0%) could resist the degradation of hydrogels. In addition, the thermodynamic properties of hydrogels also improved by the GA addition. Overall, these findings suggested that gliadin can be applied for fabricating hydrogels with tunable mechanical properties, which will unlock the high-utilization of gliadin as biopolymer and biocompatible materials.

The adaptive evolution and coevolution of the ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit ( rbcL) gene in the genus Hildenbrandia were studied based on phylogenetic tree construction and the physicochemical properties and the secondary structures of protein encoded by rbcL (Rubisco large subunit) were analyzed. The amino acids compositions and grand average of hydropathicity of freshwater H. rivularis and marine H. rubra were similar. Rubisco large subunit of Hildenbrandia was hydrophilic and the secondary structure was primarily composed of α-helixes and β-sheets, revealing the relatively stable structure of this protein. The predicted phosphorylation sites in H. rivularis and H. rubra were 33 and 36, respectively. No positive selection sites were detected in the genus Hildenbrandia, implying that rbcL gene evolved either neutrally or under purifying selection. A total of 41 coevolutionary groups were detected in the Rubisco large subunit of Hildenbrandia and the coevolving sites are in closer proximity in 3-dimensional structure of the protein. Despite the long evolutionary history, rbcL gene in genus Hildenbrandia under different environments is rather conservative.

Cold stress severely affects plant growth and yield. C-repeat binding factors (CBFs) play important roles in the response to cold stress. In the present study, we identified an R2R3-MYB transcription factor (TF) MdMYB23 from apple (Malus × domestic) using transcriptome analyses, which was notably induced in response to cold stress. Transgenic apple calli and Arabidopsis with overexpression of MdMYB23 exhibited increased cold tolerance. Electrophoretic mobility shift assay (EMSA) and transient expression assays indicated that MdMYB23 directly bound to the promoters of MdCBF1 and MdCBF2 and activated their expression. MdMYB23 interacted with the promoter of MdANR, a key modulator of proanthocyanidin biosynthesis, and activated its expression to promote proanthocyanidin accumulation and reactive oxygen species (ROS) scavenging. MdBT2 was identified as an MdMYB23-interacting protein using yeast two-hybrid (Y2H), pull-down, and bimolecular fluorescence complementation (BiFC) assays. MdBT2 repressed cold tolerance and proanthocyanidin accumulation by promoting the degradation of MdMYB23 protein. Our findings shed light on the functions of MYB TFs and underlying mechanism in the modulation of plant cold tolerance.

Objective Multiple mechanisms participated in the cell death after fluid percussion traumatic brain injury (TBI). In the present study, we evaluated the effect on cell death in the injured cortex after fluid percussion TBI and investigated a possible role of autophagy. Methods TBI model was induced by a fluid percussion TBI device. Moderate hypothermia (32°C) was achieved by partial immersion in a water bath (0°C) under general anesthesia for 4 hours. All rats were killed at 6 or 24 hours after TBI. Results Cleaved caspase 3 evaluated with Western blotting and terminal deoxynucleotidyl transferase-mediated 2′-deoxyuridine 5′-triphosphate-biotin nick end labeling-positive cells in injured cortex were significantly increased 6 hours after fluid percussion TBI and were further up-regulated 24 hours after TBI, dramatic up-regulation of Beclin-1 and protein light chain-3 expression levels was also observed. Further up-regulation of biomarkers of autophagy, attenuation of caspase 3 up-regulation and reduction of cell death was observed after 4 hours of hypothermia. Immunofluorescence analysis for cell localization demonstrated that protein light chain-3- and Beclin-1-positive cells included neurons and glial cells in the injured cortex after TBI and hypothermic treatment. By ultrastructural observation, autolysosomes in injured cortex were significantly increased at 6 and 24 hours after TBI and were further up-regulated after TBI hypothermic treatment. Conclusions These data suggest that hypothermic treatment could attenuate TBI-induced cell death in this fluid percussion TBI model, possibly through activation of autophagy pathway.

Cold stress is an adverse stimulus that affects plant growth and development, and the C-repeat binding factor (CBF) cold-regulatory cascade has been regarded as a master regulator in the plant response to cold stress. Here, we showed that a NAC transcription factor modulated low-temperature tolerance. MdNAC029/MdNAP, an apple NAC gene was isolated and its role in regulating cold tolerance was investigated. MdNAC029 was responsive to low-temperature treatment, and over-expression of MdNAC029 reduced cold tolerance in apple calli and Arabidopsis. Furthermore, EMSA assays and transient expression assays demonstrated that MdNAC029 directly repressed the expression of MdCBF1 and MdCBF4 by binding to their promoters. Taken together, our data suggest that MdNAC029 functions as a negative regulator in regulating plant cold tolerance in a CBF-dependent manner, providing a deeper understanding of NAC transcription-factor-mediated cold tolerance.

RCD1 is a member of the plant-specific SRO protein family. Several SRO genes have been functionally identified in the regulation of abiotic stresses in Arabidopsis and other plant species. However, the function of SROs is largely unknown in apple (Malus×domestica). In this study, six MdSRO-encoding genes were isolated, categorized into two types and mapped to six chromosomes. The phylogenetic analysis demonstrated that the sequences of the AtSRO and MdSRO proteins are highly conserved. Subsequently, expression analysis showed that MdSRO genes had different expression profiles in different tissues and in response to various stresses. Finally, MdRCD1 was isolated for functional identification. The results showed that resistance to oxidation stress in apple calli was enhanced by MdRCD1 overexpression and weakened by MdRCD1 suppression. MdRCD1 also played a crucial role in the regulation of ROS homeostasis in transgenic apple calli and Arabidopsis. Ectopic expression of MdRCD1 significantly enhanced resistance to salt and oxidative stresses in transgenic lines. In addition, MdRCD1 also enhanced drought tolerance due to its influence on stomatal opening. Based on these results, we conclude that MdRCD1 is an important regulator in abiotic stress response.

The basic leucine zipper (bZIP) transcription factor HY5 plays a multifaceted role in plant growth and development. Here the apple MdHY5 gene was cloned based on its homology with Arabidopsis HY5. Expression analysis demonstrated that MdHY5 transcription was induced by light and abscisic acid treatments. Electrophoretic mobility shift assays and transient expression assays subsequently showed that MdHY5 positively regulated both its own transcription and that of MdMYB10 by binding to E-box and G-box motifs, respectively. Furthermore, we obtained transgenic apple calli that overexpressed the MdHY5 gene, and apple calli coloration assays showed that MdHY5 promoted anthocyanin accumulation by regulating expression of the MdMYB10 gene and downstream anthocyanin biosynthesis genes. In addition, the transcript levels of a series of nitrate reductase genes and nitrate uptake genes in both wild-type and transgenic apple calli were detected. In association with increased nitrate reductase activities and nitrate contents, the results indicated that MdHY5 might be an important regulator in nutrient assimilation. Taken together, these results indicate that MdHY5 plays a vital role in anthocyanin accumulation and nitrate assimilation in apple. The regulatory gene HY5 plays an important role in two biochemical pathways controlling fruit quality and coloration in apple. The gene HY5 has long been recognized as a key regulator of other genes, controlling multiple processes in plant growth and development. A team from Shandong Agricultural University, China, led by Xiao-Fei Wang and Yu-Jin Hao, investigated the gene‘s action in apples, one of the world‘s most important fruit crops. They found that HY5 strongly influences the production of pigments known as anthocyanins, which help give apples their appealing color. They also found evidence that it is involved in regulating nitrogen uptake, a crucial process for plant growth, and that its action is light-activated. These results may prove useful to breeders engineering better quality and more attractive apples for the consumer market.

Cytochrome P450s play an important role in plant growth and are involved in multiple stresses response. However, little is known about the functions of cytochrome P450s in apple. Here, a Malus × domestica cytochrome P450 monooxygenase 1 gene, MdCYPM1, was identified and subsequently cloned from apple ‘Gala’ (Malus × domestica). To verify the functions of MdCYPM1, we generated transgenic Arabidopsis plants expressing the apple MdCYPM1 gene under the control of the Cauliflower mosaic virus 35S promoter. Four transgenic lines (#3, #5, #7 and #8) were selected for further study. The transgenic plants exhibited a series of skotomorphogenesis phenotypes relative to wild-type controls, such as reduction of the chlorophyll, anthocyanins content and hypocotyls elongation. In addition, overexpression of MdCYPM1 influenced auxin transport and flowering time in transgenic Arabidopsis. Furthermore, MdCYPM1 expression was induced by salt and mannitol treatments, and the transgenic plants were negatively regulated by salinity and osmotic stresses during germination. These results suggest that MdCYPM1 plays a vital role in plant growth and development.

As part of ongoing studies of the lichen family Graphidaceae in China, the status of all taxa traditionally assigned to the genus Phaeographina reported from China is resolved in the present paper. Five new combinations are proposed: Phaeographis pleiospora (Zahlbr.) Z.F. Jia & Lücking, comb. nov., Platygramme elaeoplaca (Zahlbr.) Z.F. Jia & Lücking, comb. nov., Platythecium maximum (Groenh.) Z.F. Jia & Lücking, comb. nov., P. pyrrhochroa (Mont. & Bosch) Z.F. Jia & Lücking, comb. nov., and Sarcographina heterospora (Nyl.) Z.F. Jia & Lücking, comb. nov. Six new synonyms are established: Phaeographina callospora Zahlbr. [= Diorygma hieroglyphicum (Pers.) Staiger & Kalb], P. fukiensis Zahlbr. [= Pallidogramme chrysenteron (Mont.) Staiger, Kalb & Lücking], P. fukiensis var. substriata Zahlbr. [= Pallidogramme chrysenteron (Mont.) Staiger, Kalb & Lücking], P. granulans Zahlbr. [= Platygramme platyloma (Müll. Arg.) M. Nakan. & Kashiw.], P. pluvisilvarum Zahlbr. [= Graphis alpestris (Zahlbr.) Staiger], and P. valida Zahlbr. [= Thecographa prosiliens (Mont. & Bosch) A. Massal.]. Two additional synonyms are reported: Phaeographina subrigida (Nyl.) Zahlbr. is synonymized under Platygramme platyloma (Müll. Arg.) M. Nakan. & Kashiw., and Platythecium dimorphodes (Nyl.) Staiger under Platythecium pyrrhochroum (Mont. & Bosch) Z.F. Jia & Lücking.

Objectives We aimed to investigate whether CD147 can up-regulate the chemotactic, adhesive and invasive properties of human neutrophils and to determine the mechanism underlying this process. Methods Human promyelocytic leukaemia cells (HL-60) cells and peripheral blood or synovial fluid neutrophils were isolated from RA patients. Under cyclophilin A (CypA) stimulation, chemotaxis, adhesion potential and invasion ability were assessed using chemotaxis, adhesion and invasiveness assays. Lipid raft isolation and western blot were used to determine the mechanism underlying the effects of CypA stimulation. Results CD147 up-regulates the calcium-induced chemotaxis, adhesion ability and invasiveness of human neutrophils in RA patients. Transient receptor potential melastatin 7 may be responsible for this phenomenon. Conclusion These findings suggest that in RA patients, abundant CypA up-regulates the calcium-induced chemotactic, adhesive and invasive properties of neutrophils via direct binding to CD147. Cyclophilin-CD147 interactions might contribute to the destruction of cartilage and bone in RA.